Fatty acids, C18-unsatd., dimers, oligomeric reaction products with triethylenetetramine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

28 November to 05 December 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted according to relevant testing guidelines, with no significant deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine

EC Number:

500-191-5

EC Name:

Fatty acids, C18-unsatd., dimers, oligomeric reaction products with tall-oil fatty acids and triethylenetetramine

Cas Number:

68082-29-1

IUPAC Name:

Fatty acids, C18-unsatd, dimers, polymers with tall-oil fatty acids and triethylenetetramine

Constituent 2

Reference substance name:

TOFA_DimerFA_TETA_PAA

IUPAC Name:

TOFA_DimerFA_TETA_PAA

Test material form:

liquid: viscous

Details on test material:

The test material, TOFA_DimerFA_TETA_PAA, was a pale yellow viscous material, with batch number BB 001030 V1. The test material was stored in a sealed container at room temperature, in the dark. The analytical purity was 100%.

Test animals / tissue source

Species:

other: Not applicable - in vitro study

Strain:

other: Not applicable - in vitro study

Details on test animals or tissues and environmental conditions:

Not applicable - in vitro study

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: negative and positive control substances were included

Amount / concentration applied:

750 µL. One cornea was treated with 1 mL of the test material, but this was not considered to have affected the integrity or outcome of the study.

Duration of treatment / exposure:

10 minutes

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

The test material was applied to a separate group of three corneas.

Details on study design:

Bovine eyes were supplied by a local abbatoir. The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin at 100 IU/mL and streptomycin at 100 µg/mL) in a suitably sized container and transported on the same day to the testing facility. On arrival the eyes were examined for defects including increased opacity, scratches and neovascularisation. Only corneas without such defects were used.
Upon arrival at the test facility, the corneas were excised from the eyes and loaded onto specifically designed holders. Both chambers of each holder were filled with pre-warmed Eagle’s Minimal Essential Medium (EMEM), ensuring that no bubbles were formed. The holders were incubated at 32°C ± 1°C for at least 1 hour. After the incubation, the media was removed from both the anterior and posterior chambers. Fresh media was added to the posterior chamber first and then the anterior chamber (this media replacement order ensured the cornea retained its natural curvature as much as possible). The opacity of each cornea was measured using an opacitometer. Any corneas found to have scratches or increased neovascularisation or an opacity of >7 opacity units when examined prior to treatment were discarded.
A volume of 750 µL of test material was applied to each of three corneas. The corneas were then incubated for 10 minutes at 32°C ± 1°C. After the incubation period, each cornea was washed with with media containing phenol red (as a pH indicator) until this indicator showed no pH effect occurring (and demonstrating that the test article had been removed successfully). The corneas were then washed in media without phenol red and the opacities measured. The corneas were incubated (horizontally) for 2 hours ± 10 minutes after which, the opacities were measured and then the anterior chamber emptied. For the permeability endpoint, 1 mL of sodium fluorescein (4 mg/mL solution) was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following this period, the media in the posterior chamber was removed and held in a labelled tube. Three 350 μL aliquots of this media (per cornea) were analysed for optical density at 490 nanometres (OD490) using a spectrophotometer.
Positive (dimethyl formamide) and negative (0.9% sodium chloride solution) controls were handled in an identical manner.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

10 minutes

Value:

31.9

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1. Corneal opacity

Group	Cornea number	Initial opacity	Post incubation opacity	Change in opacity	Mean change in opacity	Corrected opacity	Mean corrected opacity
Test material	3	2	51	49	N/A	48.3	30.3
	10	2	21	19		18.3
	32	6	31	25		24.3
Negative Control	15	4	4	0	4	-0.7	0.0
	4	4	5	1		0.3
	37	4	5	1		0.3
Positive Control	17	4	63	59	N/A	58.3	50.7
	23	3	52	49		48.3
	30	6	52	46		45.3

Table 2. Corneal permeability

Group	Cornea number	Mean blank OD490	OD490	Corrected OD490	Mean Corrected OD490	Final Corrected OD490	Mean group Corrected OD490
Test material	3		0.208	0.202	N/A	0.098	0.105
	10		0.065	0.059		-0.045
	32		0.372	0.366		0.262
Negative Control	15	0.06	0.058	0.052	0.006	-0.052	0.000
	4		0.150	0.143		0.040
	37		0.122	0.116		0.012
Positive Control	47		0.356	0.350	N/A	0.246	0.373
	23		0.670	0.663		0.560
	30		0.423	0.417		0.314

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The calculated IVIS score was 31.90, this is below the trigger value of 55.1 therefore the test material is not considered to be corrosive or severely irritating to the eye.

Executive summary:

The potential for TOFA_DimerFA_TETA_PAA to cause corrosion or severe irritation was evaluated in excised bovine corneas using the bovine corneal opacity and permeability (BCOP) assay, according to OECD Test Guideline 437 and GLP.

A volume of 750 µL test material was applied to three separate corneas, followed by a 10 minute incubation period at 32°C ± 1°C. Following incubation each cornea was washed with media containing phenol red, followed by media without phenol red. The opacities were then measured and the anterior chamber was emptied. To evaluate permeability, sodium fluorescein solution was added into the anterior chamber and the corneas were incubated in the vertical position for 1.5 hours ± 5 minutes. Following incubation the media in the posterior chamber was removed and three 350 µL aliquots of the media (per cornea) were analysed for optical density at 490 nm (OD₄₉₀). The negative control was 0.9% sodium chloride solution, and the positive control was dimethyl formamide.

The opacity and permeability measurements were used to calculate an In Vitro Irritancy Score (IVIS). The calculated IVIS for the test material was 31.90, this is below the trigger value of 55.1 therefore the test material is not considered to be corrosive or severely irritating to the eye.